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Increased Myoepithelial Cells of Bronchial Submucosal Glands In Asthma Thorax: first published as 10.1136/thx.2008.111435 on 8 December 2009. Downloaded from Increased myoepithelial cells of bronchial submucosal glands in fatal asthma F H Y Green,1 D J Williams,1 A James,2,3 L J McPhee,1 I Mitchell,1 T Mauad4 c Additional data are published ABSTRACT secreted into the mucous gland ducts and from online only at http://thorax.bmj. Background: Fatal asthma is characterised by enlarge- there expelled into the airway lumen. com/content/vol65/issue1 ment of bronchial mucous glands and tenacious plugs of Myoepithelial cells are ubiquitous components 1 Respiratory Research Group, mucus in the airway lumen. Myoepithelial cells, located of exocrine glands. They lie between the basement Faculty of Medicine, University within the mucous glands, contain contractile proteins membrane and the basal surface of the acinar cells of Calgary, Alberta, Canada; 2 Department of Pulmonary which provide structural support to mucous cells and and have ‘‘octopus-like’’ branching processes which 10 Physiology, West Australian actively facilitate glandular secretion. extend between the secretory epithelial cells. Sleep Disorders Research Objectives: To determine if myoepithelial cells are They have been studied in the salivary, mammary, Institute, Queen Elizabeth II increased in the bronchial submucosal glands of patients prostate, lachrymal and sweat glands in several Medical Centre, Perth, Western species.10–12 Meyrick and Reid13 described the Australia, Australia; 3 School of with fatal asthma. Medicine and Pharmacology, Methods: Autopsied lungs from 12 patients with fatal anatomical features of myoepithelial cells beneath University of Western Australia, asthma (FA), 12 patients with asthma dying of non- the serous, mucous and collecting duct cells of Perth, Western Australia, human bronchial submucosal glands. However, the 4 respiratory causes (NFA) and 12 non-asthma control Australia; Laboratory of Air mechanism of mucus expulsion from the glandular Pollution, Department of cases (NAC) were obtained through the Prairie Provinces Pathology, Sao Paulo University Asthma Study. Transverse sections of segmental bronchi acini has not been studied in human bronchial Medical School, Sao Paulo, SP, from three lobes were stained for mucus and smooth mucous glands. Brazil muscle actin and the area fractions of mucous plugs, Myoepithelial cells are contractile in nature, mucous glands and myoepithelial cells determined by possessing myofilaments composed of actin and Correspondence to: 10 13 14 point counting. The fine structure of the myoepithelial myosin. Their contraction contributes to Dr F H Y Green, Pathology and 14 Laboratory Medicine, Faculty of cells was examined by electron microscopy. glandular secretion. The mechanism of stimula- Medicine, University of Calgary, Results: FA was characterised by significant increases in tion varies by gland type and species. Contraction 3330 Hospital Drive NW, mucous gland (p = 0.003), mucous plug (p = 0.004) and may be caused by cholinergic, adrenergic or non- Calgary, Alberta T2N 4N1, 15 Canada; [email protected] myoepithelial cell areas (p = 0.017) compared with NAC. adrenergic/non-cholinergic mechanisms. When the ratio of myoepithelial cell area to total gland As the area of the submucosal glands is increased 3 5–7 Received 11 December 2008 area was examined, there was a disproportionate and in fatal asthma, it is reasonable to assume that Accepted 18 October 2009 the myoepithelial cell network is also increased in significant increase in FA compared with NAC http://thorax.bmj.com/ Published Online First relation to the increase in gland size. Given the 8 December 2009 (p = 0.014). Electron microscopy of FA cases revealed hypertrophy of the myoepithelial cells with increased striking amount of mucus found at autopsy in the airways in status asthmaticus and the marked intracellular myofilaments. The NFA group showed 47 changes in these features that were intermediate increase in airway smooth muscle, we hypothe- between the FA and NAC groups but the differences were sised that contractile elements are disproportio- not significant. nately increased within the mucous glands in fatal Conclusions: Bronchial mucous glands and mucous asthma. We report that this is the case—a finding that has important implications for understanding on September 25, 2021 by guest. Protected copyright. gland myoepithelial cell smooth muscle actin are and preventing death from asthma. increased in fatal asthma and may contribute to asphyxia due to mucous plugging. METHODS Subjects and study design The World Health Organization estimates that The study was based on autopsy materials 255 000 people died of asthma in 2005 and that 300 collected for the Prairie Provinces Asthma Study million people are currently affected by asthma.1 (PPAS), a multicentre study of asthma fatalities Death occurs by asphyxiation owing to airway occurring in Alberta, Saskatchewan and Manitoba closure through bronchoconstriction and mucous from November 1992 to October 1995.16 Cases plugging on a background of inflammation and were defined as subjects dying of asthma (fatal airway wall remodelling.2–4 Research has focused asthma, FA). There were two control groups: a more on the changes in airway smooth muscle and non-fatal asthma (NFA) group comprising subjects less on the enlarged mucous glands and tenacious with a history of asthma but who had died of non- mucous plugs.3 5–7 Overproduction of mucus in respiratory causes and a non-asthma control patients with asthma stems from a combination of (NAC) group with no history of asthma or other hyperplasia of goblet cells and enlargement of the respiratory disease at death. Deaths occurring in bronchial submucosal glands.5 The mucus in individuals with a history of asthma were reported asthma has altered viscoelastic and biochemical to the study team through the medical examiners properties that contribute to adhesivity and or coroner’s offices, hospitals and provincial vital impaired clearance.89In asthma there is an increase statistics departments. Provincial departments of in the ratio of mucous to serous cells within the vital statistics were contacted every 3 months to mucous glands.5 Mucus produced in the glands is ensure that no deaths classified as asthma in this 32 Thorax 2010;65:32–38. doi:10.1136/thx.2008.111435 Asthma Thorax: first published as 10.1136/thx.2008.111435 on 8 December 2009. Downloaded from age range were missed. NAC cases were obtained from the lumen, interstitium and blood vessels. These area fractions were Alberta Medical Examiner’s Office and local hospitals. The estimated as the number of points falling on the feature of criteria used by the pathologists and clinicians to classify the interest divided by the total number of points falling on the cases and control subjects, as well as the inclusion/exclusion mucous gland. criteria, are available in the online supplement. Further details of the morphometric methods are given in the After notification of death, the study team contacted the next online supplement. of kin to obtain consent for autopsy. The next of kin were asked to complete a questionnaire that sought information on asthma Electron microscopy severity, age of onset, duration of asthma, asthma medications Tissue for ultrastructural analysis was obtained from four and smoking history. Those with a history of asthma were subjects who had died of asthma (FA) and underwent autopsy assigned a category for asthma severity, unrelated to the cause at the Department of Pathology, Sa˜oPaulo University between of death, based on 2006 guidelines from the Global Initiative for 2005 and 2007. All had a history of asthma and no other lung 17 Asthma. Subjects were classified as having severe asthma if disease. Clinical data including treatment, smoking habits, they were using oral corticosteroids, reported hospitalisations duration of disease and previous hospitalisations were obtained for asthma (ever) or had daily symptoms. Subjects who had by administering a questionnaire to the relatives.19 Three none of the above but had symptoms on most days or nights control subjects were studied, all non-smokers with no history (more than 3 days per week), used regular inhaled corticoster- of asthma, wheeze, use of asthma medications or other lung oids or used reliever medications on most days or nights were disease and no gross or microscopic evidence of asthma at classified as having moderate asthma. All other cases were autopsy. classified as mild. Small (26262 mm) fragments of lobar bronchial wall were fixed in 2% glutaraldehyde dissolved in 0.15 M phosphate buffer Tissue sampling for light microscopy and morphometry at pH 7.2 for 1 h, followed by post-fixation in 1% osmium Left lungs taken at autopsy were fixed in inflation via the blood tetroxide dissolved in 0.9% sodium chloride for 1 h and vessels and airways with glutaraldehyde fixative (2.5% in 0.1 M embedded in Araldite resin. Ultrathin sections were studied phosphate buffer, pH 7.3). The airway pressure was with a transmission electron microscope. 20 cm H2O. The dual fixation method was developed to circumvent the poor airway perfusion resulting from mucous Statistical analysis plugs in the FA group. Transverse sections from segmental A total of 108 cases and controls had been accessioned to the bronchi were used from three sites: left upper lobe (LUL), left PPAS. A subset of 36 cases (12 FA, 12 NFA, 12 NAC) was used in lower lobe anterior bronchus (LAB) and left lower lobe posterior this study. This sample size was based on preliminary data for bronchus (LPB). Segmental bronchi were selected because they airway smooth muscle which revealed that 12 cases per group contained the greatest proportion of mucous glands relative to would provide sufficient power (77%) to detect a significant airway wall size. Tissue blocks were embedded in paraffin wax effect. Further information regarding the power calculation is http://thorax.bmj.com/ and sections stained with haematoxylin and eosin (H&E) and available in the online supplement. From the 108 subjects we Alcian blue/PAS (AB/PAS) at pH 2.5 for characterisation of randomly selected 12 subjects per group such that each group mucous plugs and mucous cells.
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